TY - JOUR
T1 - Development of a PC12 Cell Based Assay for Screening Catechol- O-methyltransferase Inhibitors
AU - Zhang, Gongliang
AU - Buchler, Ingrid P.
AU - Depasquale, Michael
AU - Wormald, Michael
AU - Liao, Gangling
AU - Wei, Huijun
AU - Barrow, James C.
AU - Carr, Gregory V.
N1 - Funding Information:
This research was funded by NIH grant R01 MH107126 and the Lieber Institute for Brain Development.
Publisher Copyright:
© 2019 American Chemical Society.
PY - 2019/10/16
Y1 - 2019/10/16
N2 - The male rat adrenal pheochromocytoma cell-derived PC12 cell line can synthesize and release catecholamine neurotransmitters, and it has been widely used as a model system in cell biology and toxicology research. Catechol-O-methyltransferase (COMT) is involved in the inactivation of the catecholamine neurotransmitters, and it is particularly important for the regulation of dopamine. In this study, we explored the feasibility of using PC12 cells as an in vitro drug screening platform to compare the activity of multiple COMT inhibitors. Incubation of PC12 cells with tolcapone, a highly potent and selective COMT inhibitor, increased the concentrations of dopamine and its metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) while reducing the metabolites 3-methoxytyramine (3-MT) and homovanillic acid (HVA) in the cell culture medium. LIBD-3, a novel, non-nitrocatechol COMT inhibitor, produced similar effects compared to tolcapone. LIBD-4, a less potent inhibitor, exhibited the expected right-shift in functional inhibition in the assay. These results match the known in vivo effects of COMT inhibition in rodents. Together, these data support the continued use of PC12 cells as an in vitro screen that bridges cell-free enzyme assays and more costly in vivo assays.
AB - The male rat adrenal pheochromocytoma cell-derived PC12 cell line can synthesize and release catecholamine neurotransmitters, and it has been widely used as a model system in cell biology and toxicology research. Catechol-O-methyltransferase (COMT) is involved in the inactivation of the catecholamine neurotransmitters, and it is particularly important for the regulation of dopamine. In this study, we explored the feasibility of using PC12 cells as an in vitro drug screening platform to compare the activity of multiple COMT inhibitors. Incubation of PC12 cells with tolcapone, a highly potent and selective COMT inhibitor, increased the concentrations of dopamine and its metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) while reducing the metabolites 3-methoxytyramine (3-MT) and homovanillic acid (HVA) in the cell culture medium. LIBD-3, a novel, non-nitrocatechol COMT inhibitor, produced similar effects compared to tolcapone. LIBD-4, a less potent inhibitor, exhibited the expected right-shift in functional inhibition in the assay. These results match the known in vivo effects of COMT inhibition in rodents. Together, these data support the continued use of PC12 cells as an in vitro screen that bridges cell-free enzyme assays and more costly in vivo assays.
KW - PC12 cells
KW - catechol-O-methyltransferase
KW - dopamine
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U2 - 10.1021/acschemneuro.9b00395
DO - 10.1021/acschemneuro.9b00395
M3 - Article
C2 - 31491076
AN - SCOPUS:85072883703
SN - 1948-7193
VL - 10
SP - 4221
EP - 4226
JO - ACS Chemical Neuroscience
JF - ACS Chemical Neuroscience
IS - 10
ER -